{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,16]],"date-time":"2025-05-16T05:48:39Z","timestamp":1747374519648,"version":"3.40.5"},"reference-count":24,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2023,4,14]],"date-time":"2023-04-14T00:00:00Z","timestamp":1681430400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sports, Science, and Technology of Japan","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Ministry of Higher Education, culture mission affairs, Egypt"},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"crossref","award":["P18091"],"award-info":[{"award-number":["P18091"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>In this study, we designed novel truncated <jats:italic><jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" ext-link-type=\"uri\" xlink:href=\"https:\/\/www.sciencedirect.com\/topics\/immunology-and-microbiology\/babesia\" xlink:type=\"simple\">Babesia<\/jats:ext-link> caballi<\/jats:italic> (<jats:italic>B<\/jats:italic>. <jats:italic>caballi<\/jats:italic>) recombinant proteins from the previously used <jats:italic>B<\/jats:italic>. <jats:italic>caballi<\/jats:italic> proteins; 134-Kilodalton Protein (rBC134) and Merozoite Rhoptry 48 Protein (rBC48). Then, we evaluated the diagnostic performance of the newly designed proteins when used as a single antigen or when used as cocktail antigen consists of rBC134 full length (rBC134f) + newly designed rBC48 (rBC48t) or newly designed rBC134 (rBC134t) + rBC48t for the detection of <jats:italic>B<\/jats:italic>. <jats:italic>caballi<\/jats:italic> infection in horse using indirect enzyme-linked immunosorbent assay (iELISA). We used one dose and a half of each antigen in the cocktail formulas. The serum samples were collected from different endemic areas in addition to the sera collected from horses experimentally infected with <jats:italic>B<\/jats:italic>. <jats:italic>caballi<\/jats:italic> were used in the present study. Cocktail antigen in full dose of (rBC134f + rBC48t) exhibited the highest <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" ext-link-type=\"uri\" xlink:href=\"https:\/\/www.sciencedirect.com\/topics\/immunology-and-microbiology\/optical-density\" xlink:type=\"simple\">optical density<\/jats:ext-link> (OD) values with <jats:italic>B<\/jats:italic>. <jats:italic>caballi<\/jats:italic>\u2013infected sera and showed the lowest OD values with normal equine sera or <jats:italic>B<\/jats:italic>. <jats:italic>caballi<\/jats:italic>, and <jats:italic>Theileria equi<\/jats:italic> mixed infected sera in comparison with the single antigen. Interestingly, the same cocktail antigen exhibited the highest concordance rate (76.74%) and kappa value (0.79) in the screening of 200 field serum samples collected from five <jats:italic>B<\/jats:italic>. <jats:italic>caballi<\/jats:italic> endemic countries, including South Africa (n = 40), Ghana (n = 40), Mongolia (n = 40), Thailand (n = 40), and China (n = 40) using iELISA and the results were compared to those of indirect fluorescent antibody test (IFAT) as a reference. Moreover, the identified promising cocktail full dose antigen (rBC134f + rBC48t) showed that it can detect the infection as early as the 4<jats:sup>th<\/jats:sup> day post-infection in sera collected from experimentally infected horses. The obtained results revealed the reliability of the rBC134f + rBC48t cocktail antigen when used in full dose for the detection of specific antibodies to <jats:italic>B<\/jats:italic>. <jats:italic>caballi<\/jats:italic> in horses which will be useful for epidemiological surveys and control of equine <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" ext-link-type=\"uri\" xlink:href=\"https:\/\/www.sciencedirect.com\/topics\/immunology-and-microbiology\/babesiosis\" xlink:type=\"simple\">babesiosis<\/jats:ext-link>.<\/jats:p>","DOI":"10.1371\/journal.pone.0284535","type":"journal-article","created":{"date-parts":[[2023,4,14]],"date-time":"2023-04-14T17:36:27Z","timestamp":1681493787000},"page":"e0284535","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":3,"title":["Development of a promising antigenic cocktail for the global detection of Babesia caballi in horse by ELISA"],"prefix":"10.1371","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9224-5731","authenticated-orcid":true,"given":"Shimaa Abd El-Salam","family":"El-Sayed","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5250-6172","authenticated-orcid":true,"given":"Mohamed Abdo","family":"Rizk","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0012-1964","authenticated-orcid":true,"given":"Hanadi B.","family":"Baghdadi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aaron Edmond","family":"Ringo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gantuya","family":"Sambuu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arifin Budiman","family":"Nugraha","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ikuo","family":"Igarashi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2023,4,14]]},"reference":[{"issue":"10","key":"pone.0284535.ref001","doi-asserted-by":"crossref","DOI":"10.3390\/ijerph16101736","article-title":"A Review on Equine Piroplasmosis: Epidemiology, Vector Ecology, Risk Factors, Host Immunity, Diagnosis and Control.","volume":"16","author":"TE Onyiche","year":"2019","journal-title":"Int J Environ Res Public Health"},{"issue":"11","key":"pone.0284535.ref002","doi-asserted-by":"crossref","DOI":"10.3390\/pathogens9110926","article-title":"Twenty Years of Equine Piroplasmosis Research: Global Distribution, Molecular Diagnosis, and Phylogeny.","volume":"9","author":"S Tirosh-Levy","year":"2020","journal-title":"Pathogens."},{"key":"pone.0284535.ref003","doi-asserted-by":"crossref","first-page":"107758","DOI":"10.1016\/j.exppara.2019.107758","article-title":"Cocktail Babesia bovis antigens for global detection of Babesia bovis infection in cattle","volume":"206","author":"S El-Sayed","year":"2019","journal-title":"Exp Parasitol"},{"key":"pone.0284535.ref004","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1146\/annurev-ento-010814-021110","article-title":"Vector ecology of equine piroplasmosis","volume":"60","author":"GA Scoles","year":"2015","journal-title":"Annu Rev Entomol"},{"issue":"4","key":"pone.0284535.ref005","doi-asserted-by":"crossref","first-page":"869","DOI":"10.1645\/GE-3064.1","article-title":"Prevalence of antibodies to Theileria equi and Babesia caballi in horses from northeastern Mexico","volume":"98","author":"MA Cantu-Martinez","year":"2012","journal-title":"J Parasitol"},{"issue":"10","key":"pone.0284535.ref006","doi-asserted-by":"crossref","first-page":"1504","DOI":"10.2174\/092986712799828355","article-title":"Current advances in detection and treatment of babesiosis","volume":"19","author":"J Mosqueda","year":"2012","journal-title":"Curr Med Chem"},{"issue":"5","key":"pone.0284535.ref007","doi-asserted-by":"crossref","first-page":"977","DOI":"10.1016\/j.apjtb.2015.09.001","article-title":"Cocktail of Theileria equi antigens for detecting infection in equines.","author":"M.A. El-Sayed SAR","year":"2015","journal-title":"Asian Pac J of Trop Biomed"},{"issue":"7","key":"pone.0284535.ref008","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1136\/jcp.44.7.569","article-title":"Comparison of IgG antibody profiles by immunoblotting in patients with acute and previous Toxoplasma gondii infection","volume":"44","author":"IL Moir","year":"1991","journal-title":"J Clin Pathol"},{"issue":"3","key":"pone.0284535.ref009","doi-asserted-by":"crossref","DOI":"10.3390\/pathogens10030270","article-title":"Development of an Indirect ELISA to Detect Equine Antibodies to Theileria haneyi.","volume":"10","author":"RG Bastos","year":"2021","journal-title":"Pathogens"},{"issue":"12","key":"pone.0284535.ref010","doi-asserted-by":"crossref","first-page":"1788","DOI":"10.1128\/CVI.00231-08","article-title":"Improved sensitivity of diagnosis of tuberculosis in patients in Korea via a cocktail enzyme-linked immunosorbent assay containing the abundantly expressed antigens of the K strain of Mycobacterium tuberculosis","volume":"15","author":"AR Shin","year":"2008","journal-title":"Clin Vaccine Immunol"},{"issue":"3","key":"pone.0284535.ref011","doi-asserted-by":"crossref","first-page":"1225","DOI":"10.1007\/s00436-015-4312-7","article-title":"Development and optimization of cocktail-ELISA for a unified surveillance of zoonotic schistosomiasis in multiple host species","volume":"114","author":"KJ Moendeg","year":"2015","journal-title":"Parasitol Res"},{"issue":"1","key":"pone.0284535.ref012","doi-asserted-by":"crossref","DOI":"10.3390\/microorganisms11010021","article-title":"Diagnostic Performance of Competitive ELISA and Western Blot Methods for the Detection of Antibodies against Theileria equi and Babesia caballi.","volume":"11","author":"G Yang","year":"2022","journal-title":"Microorganisms."},{"issue":"1","key":"pone.0284535.ref013","first-page":"211","article-title":"Molecular cloning of a Babesia caballi gene encoding the 134-kilodalton protein and evaluation of its diagnostic potential in an enzyme-linked immunosorbent assay","volume":"11","author":"Y Tamaki","year":"2004","journal-title":"Clin Diagn Lab Immunol"},{"issue":"4","key":"pone.0284535.ref014","doi-asserted-by":"crossref","first-page":"e0125276","DOI":"10.1371\/journal.pone.0125276","article-title":"Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites.","volume":"10","author":"MA Rizk","year":"2015","journal-title":"PLoS One."},{"issue":"2","key":"pone.0284535.ref015","doi-asserted-by":"crossref","first-page":"101904","DOI":"10.1016\/j.ttbdis.2022.101904","article-title":"MMV020275 and MMV020490, promising compounds from malaria box for the treatment of equine piroplasmosis.","volume":"13","author":"MA Rizk","year":"2022","journal-title":"Ticks Tick Borne Dis"},{"issue":"1","key":"pone.0284535.ref016","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1128\/JCM.42.1.359-361.2004","article-title":"Development of an immunochromatographic test with recombinant EMA-2 for the rapid detection of antibodies against Babesia equi in horses","volume":"42","author":"X Huang","year":"2004","journal-title":"J Clin Microbiol"},{"key":"pone.0284535.ref017","doi-asserted-by":"crossref","first-page":"795906","DOI":"10.3389\/fvets.2021.795906","article-title":"Identification and Characterization of P0 Protein as a Vaccine Candidate Against Babesia divergens, Blood Parasite of Veterinary and Zoonotic Importance.","volume":"8","author":"SAE El-Sayed","year":"2021","journal-title":"Front Vet Sci."},{"issue":"2","key":"pone.0284535.ref018","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1128\/CVI.00388-10","article-title":"Spherical body protein 4 is a new serological antigen for global detection of Babesia bovis infection in cattle","volume":"18","author":"MA Terkawi","year":"2011","journal-title":"Clin Vaccine Immunol"},{"issue":"6","key":"pone.0284535.ref019","doi-asserted-by":"crossref","first-page":"1334","DOI":"10.1111\/jvim.12168","article-title":"Review of equine piroplasmosis","volume":"27","author":"LN Wise","year":"2013","journal-title":"J Vet Intern Med"},{"issue":"1","key":"pone.0284535.ref020","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.vetpar.2004.08.021","article-title":"Epidemiological study of equine piroplasmosis in Mongolia","volume":"127","author":"D Boldbaatar","year":"2005","journal-title":"Vet Parasitol"},{"issue":"7","key":"pone.0284535.ref021","doi-asserted-by":"crossref","first-page":"2285","DOI":"10.1128\/JCM.37.7.2285-2290.1999","article-title":"Detection of equine antibodies to babesia caballi by recombinant B. caballi rhoptry-associated protein 1 in a competitive-inhibition enzyme-linked immunosorbent assay","volume":"37","author":"LS Kappmeyer","year":"1999","journal-title":"J Clin Microbiol"},{"issue":"11","key":"pone.0284535.ref022","doi-asserted-by":"crossref","first-page":"3475","DOI":"10.1128\/JCM.37.11.3475-3480.1999","article-title":"Cloning and expression of a 48-kilodalton Babesia caballi merozoite rhoptry protein and potential use of the recombinant antigen in an enzyme-linked immunosorbent assay","volume":"37","author":"H Ikadai","year":"1999","journal-title":"J Clin Microbiol"},{"issue":"10","key":"pone.0284535.ref023","doi-asserted-by":"crossref","first-page":"5822","DOI":"10.1128\/IAI.70.10.5822-5826.2002","article-title":"Cellular localization of Babesia bovis merozoite rhoptry-associated protein 1 and its erythrocyte-binding activity","volume":"70","author":"N Yokoyama","year":"2002","journal-title":"Infect Immun"},{"issue":"1\u20132","key":"pone.0284535.ref024","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/S0304-4017(96)01074-6","article-title":"Monoclonal antibodies against Babesia caballi and Babesia equi and their application in serodiagnosis","volume":"68","author":"A Br\u00fcning","year":"1997","journal-title":"Vet Parasitol"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0284535","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,4,14]],"date-time":"2023-04-14T17:36:55Z","timestamp":1681493815000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0284535"}},"subtitle":[],"editor":[{"given":"Etsuro","family":"Ito","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2023,4,14]]},"references-count":24,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2023,4,14]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0284535","relation":{},"ISSN":["1932-6203"],"issn-type":[{"type":"electronic","value":"1932-6203"}],"subject":[],"published":{"date-parts":[[2023,4,14]]}}}